EP1831672B1 - Mehrpunktuntersuchungsgerät - Google Patents

Mehrpunktuntersuchungsgerät Download PDF

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Publication number
EP1831672B1
EP1831672B1 EP05850861A EP05850861A EP1831672B1 EP 1831672 B1 EP1831672 B1 EP 1831672B1 EP 05850861 A EP05850861 A EP 05850861A EP 05850861 A EP05850861 A EP 05850861A EP 1831672 B1 EP1831672 B1 EP 1831672B1
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EP
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Prior art keywords
light
sample
array
spots
msg
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English (en)
French (fr)
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EP1831672A2 (de
Inventor
Derk Jan Wilfred c/o Philips I.P. & Standards GmbH KLUNDER
Maarten c/o Philips I.P. & Standards GmbH VAN HERPEN
Marcello c/o Philips I.P. & Standards GmbH BALISTRERI
Menno c/o Philips I.P. & Standards GmbH PRINS
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Koninklijke Philips NV
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Koninklijke Philips Electronics NV
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Rigid containers without fluid transport within
    • B01L3/5085—Rigid containers without fluid transport within for multiple samples, e.g. microtitration plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/06—Illumination; Optics
    • G01N2201/063—Illuminating optical parts
    • G01N2201/0635—Structured illumination, e.g. with grating

Definitions

  • the invention relates to a method and an apparatus for the investigation of a sample material with an array of light spots.
  • a laser beam is split into a plurality of excitation beams by a diffractive device.
  • the excitation beams are guided to a platform storing the sample material, where fluorescence is stimulated by an array of sample light spots. Said fluorescence is measured spatially resolved with a CCD array in order to gain information on the presence and/or amount of sample material.
  • the EP 1 283 416 A2 discloses a fluorometric imaging apparatus comprising a microlens array for generating an array of light spots in a specimen. The fluorescence that is stimulated in these light spots is measured, and input light is additionally mapped in forward direction onto a detector like a CCD camera.
  • a microscopy and lithography system that uses a microlens array for the generation of light spots is also disclosed in the WO 97/34171 .
  • the WO 94/27137 discloses an apparatus with a planar waveguide that is laterally illuminated.
  • a fluorescent sample on the surface of the waveguide is stimulated by evanescent light waves in an array of patches, and the resulting fluorescence is detected for example with a CCD array.
  • the invention comprises an apparatus for the treatment of a sample material with light.
  • the apparatus will also be called “investigation apparatus” in the following without limiting the scope of the invention.
  • sample material is to be understood in a very general sense, comprising for instance chemical elements, chemical compounds, biological material (e.g. cells), and/or mixtures thereof.
  • the apparatus comprises the following components:
  • An investigation apparatus of the aforementioned kind has two main advantages: First, the sample material in the sample chamber is investigated at a plurality of (sample) light spots simultaneously, wherein the processes take place in each spot separately. This parallelism speeds up the whole treatment procedure, allows to measure multiple analytes simultaneously, and improves the accuracy due to a better signal-to-noise ratio.
  • a second advantage is that both input light passing the sample light spots and signal light from the sample chamber are measured; thus information contained in these two measurements can be combined to find out correlations.
  • the amount of (fluorescent) signal light for example depends on both the amount of stimulating input light and the amount of fluorophores. Knowing the amount of input light at a particular sample light spot by the reference measurement therefore allows a more precise determination of the amount of fluorophores (which typically is of interest for the investigator).
  • the storage unit comprises a transparent carrier plate and a transparent cover plate which enclose the sample chamber.
  • the two plates typically consist of glass or a transparent polymer.
  • the illumination system that generates the sample light spots in the sample chamber can be realized in many different ways.
  • the illumination system first comprises a multi-spot generator (abbreviated MSG in the following) for the generation of "input light".
  • MSG multi-spot generator
  • Said input light is provided at the output side of the MSG as an array of light spots, which will be called “source light spots” in the following to distinguish them from other types of light spots.
  • the array may have a regular arrangement of source light spots, e.g. as a rectangular matrix.
  • the source light spots may particularly all have (approximately) the same shape and intensity.
  • the illumination system secondly comprises a transmission section for the transmission of input light from the source light spots of the MSG to the storage unit and, finally, to the sample light spots.
  • the signal detector that measures signal light coming from the sample chamber is preferably disposed outside the optical path of the input light. Because input light cannot directly reach the signal detector in this case, the signal light can be measured undisturbed and the signal-to-noise ratio is improved. A maximal shielding of the signal detector from the input light is achieved if it is disposed perpendicular to the propagation direction of the input light.
  • the apparatus described above may be applied for any desired kind of treatment of the sample material by light spots. Thus it may for example be used to initiate certain chemical reactions of the sample material in the sample light spots.
  • the objective is to detect, monitor and/or measure signals coming from the sample layer, particularly to measure fluorescence that was stimulated by the sample light spots.
  • the apparatus comprises the at least one signal detector which may for example be realized by photo multiplier tubes.
  • the signal detector and/or the reference detector comprise at least one array of detector elements, for example a CCD array, and an optical system for mapping the sample light spots onto said array.
  • the signal light coming from the sample light spots will be directed to different detector elements allowing a spatially resolved measurement of the sample light spots. In this way a plurality of different measurements and/or a plurality of repeated measurements of the same kind can be executed in parallel.
  • the signal light that is generated in the sample chamber propagates in all directions. Thus it may be detected in "forward direction", i.e. after traveling in the same direction as the input light propagates from the MSG to the sample light spots and beyond.
  • signal light from the sample layer may be detected in "reverse direction", i.e. a direction opposite to the propagation direction of the input light.
  • a measurement in reverse direction has the advantage that the signal light from a sample layer at the surface of the sample chamber does not have to travel largely through the sample chamber where noise might be added.
  • the measurement in reverse direction is preferable with respect to sample-handling because as there are no optics or detectors behind the sample, the sample can easily be connected to the system and there is no need for protecting the backside of the sample against e.g. dust.
  • signal light from the sample layer may be detected in "perpendicular direction", i.e. in a direction perpendicular to the propagation direction of the input light.
  • combinations of the aforementioned setups which comprise measurements in a "diagonal" direction i.e. with components in forward/reverse and perpendicular direction) are possible, too.
  • the transmission section preferably comprises a (dichroic) beam splitter that directs input light from the MSG to the storage unit and signal light from the sample chamber to the signal detector.
  • the beam splitter may particularly comprise dichroic components that show different optical behavior for different wavelengths of light, for example prisms that transmit input light of a first wavelength and simultaneously reflect fluorescence light of other wavelengths or vice versa.
  • the investigation apparatus described above allows the investigation of an area within a sample layer of the sample chamber by multiple sample light spots. In certain cases, said investigated area will not cover the whole sample layer but only a fraction thereof.
  • the apparatus is preferably adapted to shift the array of sample light spots relative to the sample chamber. This shifting may for example be achieved by a scanning unit that selectively guides light coming from the MSG or by moving the MSG (or a component of it, e.g. a mask array).
  • the apparatus is adapted to identify and re-localize positions of the sample light spots relative to the sample layer. This makes it possible to repeat a measurement at certain locations in the sample layer at least one times, thus allowing to gain additional information from a temporal development at said locations.
  • the MSG may comprise an amplitude mask, a phase mask, a holographic mask, a diffractive structure, a (micro-)lens array, a Vertical Cavity Surface-Emitting Laser (VCSEL) array and/or a multi-mode interferometer (MMI) for the generation of an array of source light spots at the output side of the MSG.
  • VCSEL Vertical Cavity Surface-Emitting Laser
  • MMI multi-mode interferometer
  • the MSG comprises a (single) light source for generating a primary light beam and an optical multiplication unit for splitting the primary light beam into an array of source light spots at the output side of the MSG.
  • the multiplication unit may for example be realized by an MMI as will be described in more detail below.
  • the splitting of a primary light beam has the advantage that only one light source (or a few light sources) is needed and the resulting source light spots have automatically the same features (wavelength, shape, intensity etc).
  • the MSG is adapted to generate an array of source light spots of coherent light, wherein said light generates a Talbot pattern during its further propagation. Due to the self-imaging character of the Talbot effect, the source light spots are periodically reproduced at certain distances, such that an image of them can be generated within the sample chamber.
  • An advantage of this application of the Talbot effect is that the transmission section requires a minimum of optical elements (lenses).
  • the MSG may particularly comprise one coherent light source.
  • the invention further comprises a method for the treatment of a sample material with light, comprising the following steps:
  • an array of source light spots of coherent light is generated from which light propagates by the Talbot effect. Due to the self-imaging character of the Talbot effect, an image of the array of source light spots may then be generated in the sample chamber with a minimum of optical elements if the sample chamber is disposed at the Talbot distance or a multiple thereof.
  • the sample light spots may particularly be generated by an array of corresponding light beams, wherein said light beams are preferably generated by splitting a primary light beam. In this way a plurality of identical light beams with required characteristics can be readily created.
  • a further development of the method is characterized in that the sample chamber is scanned with an array of sample light spots, wherein identical positions of the array are reproduced at least one times.
  • treatments can be repeated as often as desired in different locations of the sample chamber.
  • this can be used for the detection of occupied binding sites in the sample chamber, preferably for the detection of a fluorescent labeling element bound to probes in the sample chamber.
  • the method comprises in this case the scanning of the sample chamber with an array of sample light spots and the detection of target specific responses, e.g. fluorescent light, with the signal detector. If the size of the sample light spots is chosen small enough, the scanning speed is fast enough, and the concentration of binding sites is low, only one occupied binding site will be irradiated at the same time.
  • a location in the sample chamber is classified as an occupied binding site if a target specific response is observed in repeated scans of said location. Such repeated scans particularly allow to discriminate between specific and nonspecific binding.
  • fluorescence of a molecule/sample is for example used for measuring the concentration of a molecule in a solution or for detecting a bonding event (e.g. adhesion of the molecule at a layer).
  • a bonding event e.g. adhesion of the molecule at a layer.
  • a sensing array as it allows to measure multiple events, species of molecules and the location of molecules, depending on the properties of the bonding layer and the excitation light.
  • the present invention addresses this need while trying to simultaneously improve on three points: analytical performance (sensitivity, specificity, and speed), ease of use (robustness, integration), and costs.
  • FIG. 1 an exemplary setup of an investigation apparatus according to the present invention is shown.
  • Said investigation apparatus basically consists of four components or subsystems:
  • the MSG 100 may particularly comprise a (coherent) light source 101 feeding a beam of primary light into a beam splitting unit that splits or copies the primary light into an array of (identical or similar) source light spots 510 which are presented at the output side of the MSG 100.
  • the splitting unit is realized by a multi-mode interferometer MMI 106.
  • An MMI consists of a multi-modal optical waveguide. The light of the (preferably single mode) input waveguide or input spot is divided over the modes of the multi-modal waveguide section. At a given cross-section of the MMI, the intensity distribution is a periodic interference pattern between the modes of the MMI.
  • the intensity pattern at the output side of the MMI could be tuned by changing the propagation constants of the modes.
  • tuning the MMI one could also select the number of spots at the output side of the MMI and match the position of the spots with the sample layer or with optics in the transmission section 200. Because the total power in a spot is in first approximation inversely proportional to the number of spots, one could also vary/optimize the excitation power and as a consequence optimize the signal-to-noise ratio of the measurements.
  • the MMI 106 shown in Figure 1 may for example generate a one-dimensional (N ⁇ 1) array of 5 spots, with the following parameters:
  • N ⁇ M arrays of spots can be created as well. It should be remarked that the generation of the multiple spots is based on interference and can in principle be performed without significant losses.
  • Another advantage of an MMI is that it is a relatively simple method, which does not require alignment of lenses and period structures.
  • the array of source light spots 510 that is present at the output side of the MSG 100 is mapped in the transmission section 200 by collimator micro-lenses 202 and a focus (macro)-lens 204 onto sample light spots 501 in the sample layer 302 of the sample chamber 303.
  • the carrier plate 301 has the same refractive index as the focusing lens 204 in order to avoid reflections at the interface between these two components.
  • a single (macro) lens could be used instead of arrays of micro-lenses (e.g. 202) and vice versa.
  • FIG 2 shows a section through the sample chamber 303 of the apparatus of Figure 1 along the dotted line II-II.
  • Fluorescent light 500 stimulated in the sample light spots 501 propagates (also) perpendicular to the propagation direction of the input light.
  • said signal light 500 is collimated by a lens 404 and focused by a focus lens 405 onto the CCD array of the signal detector 402, where the emissions from the sample light spots 501 can be measured in a spatially resolved way.
  • Figure 3 shows a preferred way for the transmission of input light from the MSG to the sample, wherein the source light spots 510 that are present on the output side of the MSG 100 finally generate the sample light spots 501 in the sample layer 302.
  • the transmission takes place via the Talbot effect, i.e. the self-imaging of a regular pattern (in this case the array of source light spots 510) that is illuminated with a collimated beam of coherent light.
  • the array of spots 510 might also be generated by other means, for example a multi-mode interferometer (MMI), a diffractive structure, an array of (micro-)lenses or an array of VCSELs (Vertical Cavity Surface-Emitting Lasers).
  • MMI multi-mode interferometer
  • the source light spots 510 produce by interference the Talbot intensity pattern 201 which propagates through the intermediate distance into the components (glass, water) of the storage unit 300.
  • the intensity pattern of the source light spots 510 is periodically reproduced at the so-called self-imaging or Talbot distances which depend on the parameters of the setup. If for instance a grating 102 with period d is illuminated coherently, an image appears behind the grating at distances N (2 d 2 / ⁇ ), where N is an integer and ⁇ the wavelength of the light. By appropriate choice of the imaging parameters, it is thus possible to generate an image of the array of source light spots 510 at the sample side of the carrier 301.
  • N an integer and ⁇ the wavelength of the light.
  • the multiple source light spots might also be generated by a phase or holographic mask (which reproduces them roughly at 60% of the Talbot distance).
  • An important advantage of the aforementioned application of self-imaging is that it minimizes the amount of optical components like lenses in the transmission section 200, making it a simple and robust design.
  • FIG. 4 shows an embodiment for the measurement of fluorescence light in reverse direction.
  • source light spots generated by an MSG 100 are collimated and focused by lenses (not shown) at sample light spots 501 in the sample layer 302.
  • a dichroic beam splitter consisting of two prisms or wedges 206, 207 is disposed between the MSG 100 and the storage unit 300. This beam splitter has a coating such that it transmits the input light 504 and reflects the fluorescence light. Other means of separating the excitation and fluorescence light are of course not excluded from the invention.
  • Fluorescence light emitted from stimulated molecules in the sample layer 302 propagates in reverse direction (i.e. opposite to the excitation light) through the carrier plate 301 and the right wedge 207. At the inclined face of said wedge 207, the fluorescence light is reflected at right angles towards a focus lens 404 which maps it onto a CCD array 402. The fluorescence light may thus be measured separately and undisturbed from the excitation light 504.
  • the fluorescence light stimulated in the sample layer 302 can be subdivided into different components or modes according to its propagation behavior in the neighboring materials.
  • One mode that is of particular interest here is the so-called SC-mode which comprises all the fluorescence light that propagates from the sample layer 302 into the glass carrier 301 under such angles that it is totally internally reflected at the (planar) outer side of the carrier plate 301.
  • SC-mode comprises all the fluorescence light that propagates from the sample layer 302 into the glass carrier 301 under such angles that it is totally internally reflected at the (planar) outer side of the carrier plate 301.
  • a diffraction grating 305 at the outer side of the carrier 301.
  • the grating has the effect that light of the SC-modes is coupled out of the glass carrier 301 and propagates in reverse direction in light bundles 505, 506 that are highlighted in Figure 4 (light of other modes is not depicted for better clarity).
  • the light of these SC-modes is reflected at the backside of the dichroic prism 207 of the beam splitter and projected by a focus lens 404 onto a detector device 402.
  • Figure 5 schematically shows an embodiment of the investigation apparatus with a scanning unit 205 following the MSG 100 in the optical path.
  • this scanning unit 205 the array of source light spots generated by the MSG can be directed onto different sub-areas of the sample layer 302 in the storage unit 300.
  • the maximum fluorescent excitation power is limited by the saturation fluorescent intensity.
  • the measuring time can be decreased and/or the sensitivity can be increased by using the extra available laser power to apply a multi-spot approach as it is subject of the present invention. In this case the generation and scanning of the multi-spots should be done in a simple and cost effective way and preferably with no moving elements.
  • a first step to achieve a solution of the aforementioned objective comprises the use of the Talbot effect (cf. Figure 3 ), because it allows imaging of a (periodic) array of propagating spots at periodic distances without the help of lenses. In this way only the area spanned by the neighboring spots needs to be scanned for the interrogation of the total sample layer.
  • a dynamic scanning unit 205 comprising for example moving optical elements like lenses or mirrors can be used to scan the multi-spots.
  • Another possibility to move an array of multiple light spots through a sample is to scan the MSG. If for example an aperture array 102 as shown in Figure 3 is used in the MSG, the apertures only need to be moved in order to move the sample light spots 501. This is an embodiment that requires no moving lenses.
  • a characteristic feature of the investigation apparatus of Figure 5 is the single event detection with parallel spots in a scanning optical arrangement.
  • Single event detection requires a certain minimum power and energy of the emitted radiation to be detected by a sensor.
  • the choice of power conditions is elaborated in the following section.
  • the fluorophores can roughly be divided in different groups by looking to the fluorescence lifetime ⁇ fluor , the cross sections for the absorption ⁇ abs , and the fluorescent quantum efficiency ⁇ (cf. (S.W. Hell, and J. Wichmann, Opt. Lett. 19, 780, 1994),
  • a saturated-fluorescent excitation intensity I s of several ⁇ W's up to several mW's is found for a 0.2 ⁇ m 2 surface area (corresponds with an optical spot size of a DVD optical pickup unit with 0.6 NA and 650 nm).
  • the maximum applicable laser power e.g. 100 mW at the sample
  • a few (2-100) up to many (100-100000) Talbot spots can be used in parallel to scan the sensing array.
  • the fluorescent light excited by the propagating Talbot spots can be detected in the forward and the backward (reverse) propagation direction.
  • the forward fluorescent detection scheme is shown in Figure 5 .
  • the Talbot spots can be generated by different optical components, e.g. a mask with open and closed section, a multi-mode interferometer, a diffractive structure for generating an array of spots, an array of lenses or an array of VCSELs.
  • the scanning of the Talbot spots over the sample layer 302 can be obtained by scanning the multi-spot light source in the lateral direction.
  • a scanning unit 205 behind the MSG 100 allows to scan the Talbot spots.
  • the sample layer 302 of the storage unit 300 is positioned in the first Talbot plane.
  • the minimal spot size is determined by the diffraction limit.
  • a filter 405 on the other side of the storage unit 300 is used to block the red-shifted fluorescent light 500 from the input light 504 which is imaged on the pixelated reference detector 401 using an achromatic lens 403.
  • Servo signals for focusing and tracking could be generated by some spots, e.g. the four spots at the corners of the multi-spot array.
  • the reflected signal at the water interface could be used for focusing and to compensate for tilt.
  • the push-pull signal from pregrooves at the corners of the sample could be used for tracking.
  • a sample actuator with three degrees of freedom could be used to optimize the distance between the light source and the sample and the tilt between these two components.
  • the detection of the fluorescent light can be obtained in the lateral or backwards direction as in Figures 1 or 4 , respectively, because the emission is isotropic.
  • a dichroic beam splitter is required to direct the backwards fluorescent light towards the detector.
  • the length of the dichroic beam splitter is chosen such that - ignoring aberrations - the output of the beam splitter is a Talbot image of the input.
  • the input facet of the beam splitter should be in a plane where a Talbot image of the array of input spots is created and the sample side of the carrier 301 should be in a plane where a Talbot image of the output of the beam splitter is created.
  • Other configurations where the input and output facets of the beam splitter are not Talbot planes are also possible, as long as the image at the sample side of the carrier 301 is a Talbot image (ignoring aberrations) of the array of input spots.
  • the size of the dichroic beam splitter will be roughly 1 mm for a sensing array with a size 1 ⁇ 1 mm 2 .
  • the distance to the first Talbot plane (in air) for a spot pitch of 20 ⁇ m and a wavelength of 500 nm is 1.6 mm.
  • the 1 ⁇ 1 mm 2 sensing array would be simultaneously scanned by 50 ⁇ 50 Talbot spots.

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Claims (14)

  1. Vorrichtung zur Behandlung eines Probenmaterials, mit
    a) einer Speichereinheit (300), welche den Durchgang von Eingangslicht (504) erlaubt und eine Probenkammer (303) umfasst, die mit dem Probenmaterial versehen werden kann;
    b) einem Beleuchtungssystem (100, 200), um mit Eingangslicht (504) ein Array von Proben-Lichtpunkten (501) in der Probenkammer (303) zu erzeugen;
    c) einem Referenzdetektor (401) zum Messen von Eingangslicht (504), welches sich über die Proben-Lichtpunkte (501) hinaus ausbreitet;
    d) einem Signaldetektor (402) zum Messen von Signallicht (500), welches in der Probenkammer (303) erzeugt wird;
    e) Mitteln zur Referenzierung der gemessenen Signallichtmenge auf die gemessene Eingangslichtmenge für die Proben-Lichtpunkte.
  2. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass die Speichereinheit (300) eine lichtdurchlässige Trägerplatte (301) und eine lichtdurchlässige Deckplatte (304) umfasst, welche die Probenkammer (303) umgeben.
  3. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass das Beleuchtungssystem einen Mehrpunktgenerator MSG (100) zur Erzeugung von Eingangslicht in der Form eines Arrays von Lichtpunkten (510) einer Lichtquelle an seiner Ausgangsseite sowie einen Durchlässigkeitsteil (200) zum Durchlassen von Eingangslicht (504) von den Lichtpunkten (510) einer Lichtquelle zu den Proben-Lichtpunkten (501) umfasst.
  4. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass der Signaldetektor (402) außerhalb des optischen Wegs des Eingangslichts (504), vorzugsweise senkrecht zu der Ausbreitungsrichtung des Eingangslichts (504), angeordnet ist.
  5. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass der Referenzdetektor (401) und/oder der Signaldetektor (402) ein Array von Detektorelementen, im Besonderen ein CCD-Array, sowie ein optisches System (403, 404, 405, 406) umfasst, um die Proben-Lichtpunkte (501) auf dem Array abzubilden.
  6. Vorrichtung nach Anspruch 3, dadurch gekennzeichnet, dass der Durchlässigkeitsteil (200) einen Strahlteiler umfasst, der Eingangslicht (504) von dem MSG (100) zu der Speichereinheit (300) und Signallicht von der Probenkammer (303) zu dem Signaldetektor (402) leitet.
  7. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass diese das Array von Proben-Lichtpunkten (501) relativ zu der Probenkammer (303), vorzugsweise mit Hilfe einer Abtasteinheit (205), verschieben kann, um von dem MSG (100) erzeugtes Eingangslicht selektiv zu leiten.
  8. Vorrichtung nach Anspruch 1, dadurch gekennzeichnet, dass auf mindestens einer Außenfläche der Speichereinheit (300) diffraktive Strukturen (305) vorgesehen sind, die Signallicht (505, 506), welches ohne solche Strukturen innen total reflektiert würde, auskoppeln können.
  9. Vorrichtung nach Anspruch 3, dadurch gekennzeichnet, dass der MSG (100) eine Amplitudenmaske (102), eine Phasenmaske, eine holographische Maske, eine diffraktive Struktur, ein Mikrolinsenarray, ein VCSEL-Array und/oder ein Mehrmoden-Interferometer (106) zur Erzeugung eines Arrays von Lichtpunkten (510) einer Lichtquelle an der Ausgangsseite des MSGs (100) umfasst.
  10. Vorrichtung nach Anspruch 3, dadurch gekennzeichnet, dass der MSG (100) eine Lichtquelle (101) zur Erzeugung eines Primärlichtstrahls sowie eine optische Multiplikationseinheit, im Besonderen ein Mehrmoden-Interferometer (106), umfasst, um den Primärlichtstrahl in ein Array von Lichtpunkten (510) der Lichtquelle an der Ausgangsseite des MSGs (100) aufzuspalten.
  11. Vorrichtung nach Anspruch 3, dadurch gekennzeichnet, dass der MSG (100) ein Array von Lichtquellen-Lichtpunkten (510) aus kohärentem Licht erzeugen kann, die eine Talbotstruktur (201) bilden.
  12. Verfahren zur Behandlung eines Probenmaterials mit Licht, welches die folgenden Schritte umfasst:
    a) Erzeugen von Eingangslicht (504) und eines Arrays von einer Vielzahl von Proben-Lichtpunkten (501) in einer das Probenmaterial enthaltenden Probenkammer (303) mit Hilfe des Eingangslichts;
    b) Messen von Eingangslicht (504), welches sich über die Proben-Lichtpunkte (501) hinaus ausbreitet;
    c) Messen des Signallichts (500), welches in der Probenkammer (303) erzeugt wird; sowie
    d) Referenzieren der gemessenen Signallichtmenge auf die gemessene Eingangslichtquelle für die Proben-Lichtpunkte.
  13. Verfahren nach Anspruch 12, dadurch gekennzeichnet, dass das Eingangslicht (504) die Fluoreszenz (500) des Probenmaterials anregt.
  14. Verfahren nach Anspruch 12, dadurch gekennzeichnet, dass das Signallicht (500) außerhalb des optischen Wegs des Eingangslichts (504) gemessen wird.
EP05850861A 2004-12-17 2005-12-13 Mehrpunktuntersuchungsgerät Expired - Lifetime EP1831672B1 (de)

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EP05850861A EP1831672B1 (de) 2004-12-17 2005-12-13 Mehrpunktuntersuchungsgerät
PCT/IB2005/054207 WO2006064465A2 (en) 2004-12-17 2005-12-13 Multi-spot investigation apparatus

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EP1831672B1 true EP1831672B1 (de) 2008-10-22

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CN101080627A (zh) 2007-11-28
CN101080627B (zh) 2010-06-23
EP1831672A2 (de) 2007-09-12
WO2006064465A3 (en) 2006-09-14
DE602005010611D1 (de) 2008-12-04
WO2006064465A2 (en) 2006-06-22
ATE412170T1 (de) 2008-11-15
JP2008524578A (ja) 2008-07-10
US20100051788A1 (en) 2010-03-04

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